data_16457 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; The Structural Basis for the Ligand Specificity of Family 2 Carbohydrate-binding Modules ; _BMRB_accession_number 16457 _BMRB_flat_file_name bmr16457.str _Entry_type original _Submission_date 2009-08-19 _Accession_date 2009-08-19 _Entry_origination author _NMR_STAR_version 2.1.1 _Experimental_method NMR _Details . loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Simpson Peter J. . 2 Xie Hefang . . 3 Bolam David N. . 4 Gilbert Harry J. . 5 Williamson Michael P. . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count binding_constants 1 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2010-03-08 original author . stop_ _Original_release_date 2010-03-08 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title 'The Structurl Basis for the Ligand Specificity of Family 2 Carbohydrate-binding Modules' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 10973978 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Simpson Peter J. . 2 Xie Hefang . . 3 Bolam David N. . 4 Gilbert Harry J. . 5 Williamson Michael P. . stop_ _Journal_abbreviation 'J. Biol. Chem.' _Journal_name_full 'Journal of Biological Chemistry' _Journal_volume 275 _Journal_issue 52 _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 41137 _Page_last 41142 _Year 2000 _Details . save_ ################################## # Molecular system description # ################################## save_assembly _Saveframe_category molecular_system _Mol_system_name CBM2b/cellohexaose _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label 'CBM2b R262G' $CBM2b_R262G cellohexaose $entity_CE6 stop_ _System_molecular_weight . _System_physical_state native _System_oligomer_state ? _System_paramagnetic no _System_thiol_state . _Database_query_date . _Details . save_ ######################## # Monomeric polymers # ######################## save_CBM2b_R262G _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common CBM2b_R262G _Molecular_mass . _Mol_thiol_state 'not reported' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 84 _Mol_residue_sequence ; STGCSVTATRAEEWSDGFNV TYSVSGSSAWTVNLALNGSQ TIQASWNANVTGSGSTRTVT PNGSGNTVMKNGSSTTPAAT CAGS ; loop_ _Residue_seq_code _Residue_label 1 SER 2 THR 3 GLY 4 CYS 5 SER 6 VAL 7 THR 8 ALA 9 THR 10 ARG 11 ALA 12 GLU 13 GLU 14 TRP 15 SER 16 ASP 17 GLY 18 PHE 19 ASN 20 VAL 21 THR 22 TYR 23 SER 24 VAL 25 SER 26 GLY 27 SER 28 SER 29 ALA 30 TRP 31 THR 32 VAL 33 ASN 34 LEU 35 ALA 36 LEU 37 ASN 38 GLY 39 SER 40 GLN 41 THR 42 ILE 43 GLN 44 ALA 45 SER 46 TRP 47 ASN 48 ALA 49 ASN 50 VAL 51 THR 52 GLY 53 SER 54 GLY 55 SER 56 THR 57 ARG 58 THR 59 VAL 60 THR 61 PRO 62 ASN 63 GLY 64 SER 65 GLY 66 ASN 67 THR 68 VAL 69 MET 70 LYS 71 ASN 72 GLY 73 SER 74 SER 75 THR 76 THR 77 PRO 78 ALA 79 ALA 80 THR 81 CYS 82 ALA 83 GLY 84 SER stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date . save_ ############# # Ligands # ############# save_CE6 _Saveframe_category ligand _Mol_type "non-polymer (NON-POLYMER)" _Name_common CELLOHEXAOSE _BMRB_code CE6 _PDB_code CE6 _Molecular_mass 990.859 _Mol_charge 0 _Mol_paramagnetic . _Mol_aromatic no _Details . loop_ _Atom_name _PDB_atom_name _Atom_type _Atom_chirality _Atom_charge _Atom_oxidation_number _Atom_unpaired_electrons C1A C1A C . 0 . ? C2A C2A C . 0 . ? C3A C3A C . 0 . ? C4A C4A C . 0 . ? C5A C5A C . 0 . ? C6A C6A C . 0 . ? O2A O2A O . 0 . ? O3A O3A O . 0 . ? O4A O4A O . 0 . ? O5A O5A O . 0 . ? O6A O6A O . 0 . ? C1B C1B C . 0 . ? C2B C2B C . 0 . ? C3B C3B C . 0 . ? C4B C4B C . 0 . ? C5B C5B C . 0 . ? C6B C6B C . 0 . ? O2B O2B O . 0 . ? O3B O3B O . 0 . ? O4B O4B O . 0 . ? O5B O5B O . 0 . ? O6B O6B O . 0 . ? C1C C1C C . 0 . ? C2C C2C C . 0 . ? C3C C3C C . 0 . ? C4C C4C C . 0 . ? C5C C5C C . 0 . ? C6C C6C C . 0 . ? O2C O2C O . 0 . ? O3C O3C O . 0 . ? O4C O4C O . 0 . ? O5C O5C O . 0 . ? O6C O6C O . 0 . ? C1D C1D C . 0 . ? C2D C2D C . 0 . ? C3D C3D C . 0 . ? C4D C4D C . 0 . ? C5D C5D C . 0 . ? C6D C6D C . 0 . ? O2D O2D O . 0 . ? O3D O3D O . 0 . ? O4D O4D O . 0 . ? O5D O5D O . 0 . ? O6D O6D O . 0 . ? O2F O2F O . 0 . ? C2F C2F C . 0 . ? C3F C3F C . 0 . ? O3F O3F O . 0 . ? C4F C4F C . 0 . ? O4F O4F O . 0 . ? C5F C5F C . 0 . ? C6F C6F C . 0 . ? O6F O6F O . 0 . ? O5F O5F O . 0 . ? C1F C1F C . 0 . ? O4E O4E O . 0 . ? C4E C4E C . 0 . ? C5E C5E C . 0 . ? O5E O5E O . 0 . ? C6E C6E C . 0 . ? O6E O6E O . 0 . ? C3E C3E C . 0 . ? O3E O3E O . 0 . ? C2E C2E C . 0 . ? O2E O2E O . 0 . ? C1E C1E C . 0 . ? O1A O1A O . 0 . ? H1A H1A H . 0 . ? H2A H2A H . 0 . ? H3A H3A H . 0 . ? H4A H4A H . 0 . ? H5A H5A H . 0 . ? H6A1 H6A1 H . 0 . ? H6A2 H6A2 H . 0 . ? HO2A HO2A H . 0 . ? HO3A HO3A H . 0 . ? HO6A HO6A H . 0 . ? H1B H1B H . 0 . ? H2B H2B H . 0 . ? H3B H3B H . 0 . ? H4B H4B H . 0 . ? H5B H5B H . 0 . ? H6B1 H6B1 H . 0 . ? H6B2 H6B2 H . 0 . ? HO2B HO2B H . 0 . ? HO3B HO3B H . 0 . ? HO6B HO6B H . 0 . ? H1C H1C H . 0 . ? H2C H2C H . 0 . ? H3C H3C H . 0 . ? H4C H4C H . 0 . ? H5C H5C H . 0 . ? H6C1 H6C1 H . 0 . ? H6C2 H6C2 H . 0 . ? HO2C HO2C H . 0 . ? HO3C HO3C H . 0 . ? HO6C HO6C H . 0 . ? H1D H1D H . 0 . ? H2D H2D H . 0 . ? H3D H3D H . 0 . ? H4D H4D H . 0 . ? H5D H5D H . 0 . ? H6D1 H6D1 H . 0 . ? H6D2 H6D2 H . 0 . ? HO2D HO2D H . 0 . ? HO3D HO3D H . 0 . ? HO6D HO6D H . 0 . ? HO2F HO2F H . 0 . ? H2F H2F H . 0 . ? H3F H3F H . 0 . ? HO3F HO3F H . 0 . ? H4F H4F H . 0 . ? HO4F HO4F H . 0 . ? H5F H5F H . 0 . ? H6F1 H6F1 H . 0 . ? H6F2 H6F2 H . 0 . ? HO6F HO6F H . 0 . ? H1F H1F H . 0 . ? H4E H4E H . 0 . ? H5E H5E H . 0 . ? H6E1 H6E1 H . 0 . ? H6E2 H6E2 H . 0 . ? HO6E HO6E H . 0 . ? H3E H3E H . 0 . ? HO3E HO3E H . 0 . ? H2E H2E H . 0 . ? HO2E HO2E H . 0 . ? H1E H1E H . 0 . ? HO1A HO1A H . 0 . ? stop_ loop_ _Bond_order _Bond_atom_one_atom_name _Bond_atom_two_atom_name _PDB_bond_atom_one_atom_name _PDB_bond_atom_two_atom_name SING C1A C2A ? ? SING C1A O5A ? ? SING C1A O1A ? ? SING C1A H1A ? ? SING C2A C3A ? ? SING C2A O2A ? ? SING C2A H2A ? ? SING C3A O3A ? ? SING C3A C4A ? ? SING C3A H3A ? ? SING C4A O4A ? ? SING C4A C5A ? ? SING C4A H4A ? ? SING C5A C6A ? ? SING C5A O5A ? ? SING C5A H5A ? ? SING C6A O6A ? ? SING C6A H6A1 ? ? SING C6A H6A2 ? ? SING O2A HO2A ? ? SING O3A HO3A ? ? SING O4A C1B ? ? SING O6A HO6A ? ? SING C1B C2B ? ? SING C1B O5B ? ? SING C1B H1B ? ? SING C2B C3B ? ? SING C2B O2B ? ? SING C2B H2B ? ? SING C3B O3B ? ? SING C3B C4B ? ? SING C3B H3B ? ? SING C4B O4B ? ? SING C4B C5B ? ? SING C4B H4B ? ? SING C5B C6B ? ? SING C5B O5B ? ? SING C5B H5B ? ? SING C6B O6B ? ? SING C6B H6B1 ? ? SING C6B H6B2 ? ? SING O2B HO2B ? ? SING O3B HO3B ? ? SING O4B C1C ? ? SING O6B HO6B ? ? SING C1C C2C ? ? SING C1C O5C ? ? SING C1C H1C ? ? SING C2C C3C ? ? SING C2C O2C ? ? SING C2C H2C ? ? SING C3C O3C ? ? SING C3C C4C ? ? SING C3C H3C ? ? SING C4C O4C ? ? SING C4C C5C ? ? SING C4C H4C ? ? SING C5C C6C ? ? SING C5C O5C ? ? SING C5C H5C ? ? SING C6C O6C ? ? SING C6C H6C1 ? ? SING C6C H6C2 ? ? SING O2C HO2C ? ? SING O3C HO3C ? ? SING O4C C1D ? ? SING O6C HO6C ? ? SING C1D O5D ? ? SING C1D C2D ? ? SING C1D H1D ? ? SING C2D C3D ? ? SING C2D O2D ? ? SING C2D H2D ? ? SING C3D C4D ? ? SING C3D O3D ? ? SING C3D H3D ? ? SING C4D O4D ? ? SING C4D C5D ? ? SING C4D H4D ? ? SING C5D C6D ? ? SING C5D O5D ? ? SING C5D H5D ? ? SING C6D O6D ? ? SING C6D H6D1 ? ? SING C6D H6D2 ? ? SING O2D HO2D ? ? SING O3D HO3D ? ? SING O4D C1E ? ? SING O6D HO6D ? ? SING O2F C2F ? ? SING O2F HO2F ? ? SING C2F C3F ? ? SING C2F C1F ? ? SING C2F H2F ? ? SING C3F C4F ? ? SING C3F O3F ? ? SING C3F H3F ? ? SING O3F HO3F ? ? SING C4F O4F ? ? SING C4F C5F ? ? SING C4F H4F ? ? SING O4F HO4F ? ? SING C5F C6F ? ? SING C5F O5F ? ? SING C5F H5F ? ? SING C6F O6F ? ? SING C6F H6F1 ? ? SING C6F H6F2 ? ? SING O6F HO6F ? ? SING O5F C1F ? ? SING C1F O4E ? ? SING C1F H1F ? ? SING O4E C4E ? ? SING C4E C3E ? ? SING C4E C5E ? ? SING C4E H4E ? ? SING C5E C6E ? ? SING C5E O5E ? ? SING C5E H5E ? ? SING O5E C1E ? ? SING C6E O6E ? ? SING C6E H6E1 ? ? SING C6E H6E2 ? ? SING O6E HO6E ? ? SING C3E O3E ? ? SING C3E C2E ? ? SING C3E H3E ? ? SING O3E HO3E ? ? SING C2E O2E ? ? SING C2E C1E ? ? SING C2E H2E ? ? SING O2E HO2E ? ? SING C1E H1E ? ? SING O1A HO1A ? ? stop_ _Mol_thiol_state . _Sequence_homology_query_date . save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species $CBM2b_R262G 'Cellulomonas fimi' 1708 Bacteria . Cellulomonas fimi stop_ save_ ######################### # Experimental source # ######################### save_experimental_source _Saveframe_category experimental_source loop_ _Mol_label _Production_method _Host_organism_name_common _Genus _Species _Strain _Variant _Vector_name $CBM2b_R262G 'recombinant technology' . Escherichia coli BL21 DE3 pET16b stop_ save_ ##################################### # Sample contents and methodology # ##################################### ######################## # Sample description # ######################## save_sample_1 _Saveframe_category sample _Sample_type solution _Details . loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $CBM2b_R262G 100 uM [U-15N] 'sodium phosphate buffer' 50 mM 'natural abundance' D2O 10 % 'natural abundance' H2O 90 % 'natural abundance' stop_ save_ save_sample_2 _Saveframe_category sample _Sample_type solution _Details 'concentrations of ligand taken from graph' loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $CBM2b_R262G 100 uM [U-15N] $entity_CE6 50 uM 'natural abundance' 'sodium phosphate buffer' 50 mM 'natural abundance' D2O 10 % 'natural abundance' H2O 90 % 'natural abundance' stop_ save_ save_sample_3 _Saveframe_category sample _Sample_type solution _Details 'concentrations of ligand taken from graph' loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $CBM2b_R262G 100 uM [U-15N] $entity_CE6 200 uM 'natural abundance' 'sodium phosphate buffer' 50 mM 'natural abundance' D2O 10 % 'natural abundance' H2O 90 % 'natural abundance' stop_ save_ save_sample_4 _Saveframe_category sample _Sample_type solution _Details 'concentrations of ligand taken from graph' loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $CBM2b_R262G 100 uM [U-15N] $entity_CE6 800 uM 'natural abundance' 'sodium phosphate buffer' 50 mM 'natural abundance' D2O 10 % 'natural abundance' H2O 90 % 'natural abundance' stop_ save_ save_sample_5 _Saveframe_category sample _Sample_type solution _Details 'concentrations of ligand taken from graph' loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $CBM2b_R262G 100 uM [U-15N] $entity_CE6 2 mM 'natural abundance' 'sodium phosphate buffer' 50 mM 'natural abundance' D2O 10 % 'natural abundance' H2O 90 % 'natural abundance' stop_ save_ save_sample_6 _Saveframe_category sample _Sample_type solution _Details 'concentrations of ligand taken from graph' loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $CBM2b_R262G 100 uM [U-15N] $entity_CE6 4 mM 'natural abundance' 'sodium phosphate buffer' 50 mM 'natural abundance' D2O 10 % 'natural abundance' H2O 90 % 'natural abundance' stop_ save_ save_sample_7 _Saveframe_category sample _Sample_type solution _Details 'concentrations of ligand taken from graph' loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $CBM2b_R262G 100 uM [U-15N] $entity_CE6 9.5 mM 'natural abundance' 'sodium phosphate buffer' 50 mM 'natural abundance' D2O 10 % 'natural abundance' H2O 90 % 'natural abundance' stop_ save_ ############################ # Computer software used # ############################ save_EXCEL _Saveframe_category software _Name EXCEL _Version 5.0 loop_ _Vendor _Address _Electronic_address '(Microsoft Corporation)' . . stop_ loop_ _Task 'data fitting' stop_ _Details . save_ save_X-PLOR _Saveframe_category software _Name X-PLOR _Version . loop_ _Vendor _Address _Electronic_address Brunger . . stop_ loop_ _Task 'structure solution' stop_ _Details . save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_spectrometer_1 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model DRX _Field_strength 800 _Details . save_ ############################# # NMR applied experiments # ############################# save_2D_1H-15N_HSQC_1 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _Sample_label $sample_1 save_ save_2D_1H-15N_HSQC_2 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _Sample_label $sample_2 save_ save_2D_1H-15N_HSQC_3 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _Sample_label $sample_3 save_ save_2D_1H-15N_HSQC_4 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _Sample_label $sample_4 save_ save_2D_1H-15N_HSQC_5 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _Sample_label $sample_5 save_ save_2D_1H-15N_HSQC_6 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _Sample_label $sample_6 save_ save_2D_1H-15N_HSQC_7 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _Sample_label $sample_7 save_ ####################### # Sample conditions # ####################### save_sample_conditions_1 _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units 'ionic strength' 50 . mM pH 7.0 . pH pressure 1 . atm temperature 303 . K stop_ save_